Coupled vs uncoupled in vitro transcription/translation

Wade Harper jharper at bcm.tmc.edu
Thu Dec 2 21:49:43 EST 1993


In article <9311251710.AA31423 at bono.oci.utoronto.ca>,
jwoodget at ocicl.oci.utoronto.ca (Jim Woodgett) wrote:
> 
> Has anyone had good/bad experiences with Promega's TnT coupled 
> transcription/translation system?  We've always made RNA first then 
> translated in retic. lysates but this does take more time than the coupled 
> methods.  Any comparisons between the two apart from cost?
> 
> 
> 
> Jim Woodgett      Fax (416) 926 6529     Tel (416) 924 0671
> jwoodget at ocicl.oci.utoronto.ca
> _____________________________________________________________


In our hands, TnT works better than the two step method.  There are far
fewer 
internal initiation and premature termination products especially for large
proteins.  I guess that is in part because the RNA isnot nicked before
it enters the ribosome.

Wade Harper
jharper at bcm.tmc.edu



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